Kit for rapid detection and genotyping of Mycoplasma pneumoniae

Abstract

The invention provides a kit for rapid detection and genotyping of Mycoplasma pneumoniae, which belongs to the technical field of microbial detection. The kit contains 2 pairs of specific primer pairs and 2 probes used in conjunction with the primer pairs. The nucleotide sequences are as follows: As shown in SEQ ID No.1-6, the dual fluorescent quantitative PCR method is used to realize rapid detection and genotyping of Mycoplasma pneumoniae, the detection sensitivity is 3 CFU, and the accuracy of genotyping is 100% consistent with the commonly used genotyping methods , the sensitivity of typing is much higher than that of commonly used culture-dependent typing techniques, which greatly shortens the typing time, and enables genotyping while detecting Mycoplasma pneumoniae in samples, overcoming the current typing method of Mycoplasma pneumoniae It has the disadvantages of low sensitivity, laborious work and high cost, but it has good detection and typing capabilities of actual specimens and is suitable for popularization and application.

Description

technical field [0001] The invention relates to the technical field of microorganism detection, in particular to a kit for rapid detection and genotyping of Mycoplasma pneumoniae. Background technique [0002] Mycoplasma pneumoniae (Mycoplasma pneumoniae, Mp) is one of the important pathogens causing human respiratory tract infection, and about 10%-40% of community-acquired pneumonia is caused by its infection every year. Fluorescent PCR detection technology has gradually become the gold standard for the detection of Mycoplasma pneumoniae infection by virtue of its advantages of rapidity, high sensitivity and specificity, and has been widely used in clinical testing and scientific research. Genotyping Mycoplasma pneumoniae on the basis of detection is an important monitoring method for in-depth understanding of its antigenic variation and early warning and prediction of strain epidemic outbreaks. Studies have found that Mycoplasma pneumoniae can be divided into two types of...

AI Technical Summary

Problems solved by technology

[0003] Although the commonly used Mycoplasma pneumoniae typing techniques reported above have been widely used, they have obvious defects: these methods all rely on the isolation and culture of Mycoplasma pneumoniae, that is, after the Mycoplasma pneumoniae strains are isolated from the specimens, they are purified and purified. Genotyping can only be performed after culturing and extracting high-concentration Mycoplasma pneumoniae genomic DNA, and the amount of Mycoplasma pneumoniae nucleic acid in ordinary clinical specimens is not enough to support the detection by the above-mentioned typing method

This situation has led to a serious disconnection between the detection technology and the typing technology of Mycoplasma pneumoniae: after nucleic acid extraction, the rapid detection of Mycoplasma pneumoniae is carried out by fluorescent PCR technology (the test result can be obtained within 1 hour), but the typing needs to be determined first. The specimens were isolated and cultured for Mycoplasma pneumoniae strains, and the positive specimens could be tested by common typing methods after nucleic acid extraction (the typing results could be obtained after an average of 2 weeks), and the culture of Mycoplasma pneumoniae specimens was very difficult and the positive rate was low, resulting in Many positive specimens detected by fluorescent PCR cannot be isolated and cultured for strains and cannot be tested for typing, which makes the typing results biased to a certain extent

The inconsistency between the detection sensitivity of Mycoplasma pneumoniae and genotyping detection, and the asynchronous detection time limits have seriously restricted the development of Mycoplasma pneumoniae research.

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